OLIGORIBONUCLEOTIDES CONTAINING 2',5'-PHOSPHODIESTER LINKAGES EXHIBIT BINDING SELECTIVITY FOR 3',5'-RNA OVER 3',5'-SSDNA
OLIGORIBONUCLEOTIDES CONTAINING 2',5'-PHOSPHODIESTER LINKAGES EXHIBIT BINDING SELECTIVITY FOR 3',5'-RNA OVER 3',5'-SSDNA
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DOI：
10.1093/nar/21.20.4742
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发表时间：
1993-10-11
影响因子：
14.9
通讯作者：
DAMHA, MJ
中科院分区：
文献类型：
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作者：
GIANNARIS, PA;DAMHA, MJ
含有2 '，5'-磷酸二酯键的寡聚核糖核苷酸已经通过“甲硅烷基-亚磷酰胺”方法在固体载体上合成。已经使用寡腺苷酸和包括硫代磷酸酯骨架的混合碱基序列的各种寡核苷酸研究了这些寡核苷酸和互补3 '，5'-RNA链之间形成的复合物的稳定性。在许多情况下，特别是对于2 '，5'-连接的腺苷酸，UV解链曲线相当尖锐，并表现出大的增色变化。用寡聚体内的2 '，5'-键取代几个3 '，5'-键降低了复合物的T（m），并且不稳定的程度取决于相邻残基和相邻键。在本研究中制备的2 '，5'-连接的寡核糖核苷酸对互补单链RNA表现出显著的选择性超过DNA。例如，在0.01 M磷酸盐缓冲液-0.10 M NaCl（pH 7.0）中，未观察到2 '，5'-r（CCC UCU CCC UUC U）与其沃森-克里克DNA互补序列3 '，5'-d（AGAAGGGAGGG）之间的缔合。然而. 2 '，5'-r（CCC UCU CCC UUC U）与其RNA互补物3 '，5'-r（AGAAGGGAGGG）形成双链体，其在40 ℃解链。十聚体2 '，5'-r（Ap）9A与聚dT和聚rU形成复合物，但复合物[2 '，5'-r（Ap）9A ]：[聚dT]不稳定（T（m），-1 ℃），并且仅在高盐浓度下可见。鉴于它们的非天然特性和对单链RNA的显著选择性，2 '，5'-寡聚RNA及其衍生物可用作病毒mRNA翻译的选择性抑制剂。和作为亲和配体用于纯化细胞RNA。
Oligoribonucleotides containing 2',5'-phosphodiester linkages have been synthesized on a solid support by the 'silyl-phosphoramidite' method. The stability of complexes formed between these oligonucleotides and complementary 3',5'-RNA strands have been studied using oligoadenylates and a variety of oligonucleotides of mixed base sequences including phosphorothioate backbones. In many cases, particularly for 2',5'-linked adenylates, the UV melting profiles are quite sharp and exhibit large hyperchromic changes. Substituting a few 3',5'-linkages with the 2',5'-linkage within an oligomer lowers the T(m) of the complex and the degree of destabilization depends on the neighboring residues and neighboring linkages. The 2',5'-linked oligoribonucleotides prepared in this study exhibited remarkable selectivity for complementary single stranded RNA over DNA. For example, in 0.01 M phosphate buffer - 0.10 M NaCl (pH 7.0), no association was observed between 2',5'-r(CCC UCU CCC UUC U) and its Watson - Crick DNA complement 3',5'-d(AGAAGGGAGAGGG). However. 2',5'-r(CCC UCU CCC UUC U) with its RNA complement 3',5'-r(AGAAGGGAGAGGG) forms a duplex which melts at 40-degrees-C. The decamer 2',5'-r(Ap)9A forms a complex with both poly dT and poly rU but the complex [2',5'-r(Ap)9A ]: [poly dT] is unstable (T(m), - 1-degrees-C) and is seen only at high salt concentrations. In view of their unnatural character and remarkable selectivity for single stranded RNA, 2',5'-oligo-RNAs and their derivatives may find use as selective inhibitors of viral mRNA translation. and as affinity ligands for the purification of cellular RNA.